scholarly journals Characterization of impulse propagation at the microscopic level across geometrically defined expansions of excitable tissue: multiple site optical recording of transmembrane voltage (MSORTV) in patterned growth heart cell cultures.

1994 ◽  
Vol 104 (2) ◽  
pp. 287-309 ◽  
Author(s):  
S Rohr ◽  
B M Salzberg

Impulse propagation across sudden expansions of excitable tissue has been shown to exhibit various forms of conduction disturbance on a macroscopic scale, ranging from small delays to unidirectional or complete conduction block. With the present study, we attempted to characterize systematically the dependence of impulse propagation on the geometry of the underlying excitable tissue on a microscopic scale by investigating the spatio-temporal pattern of transmembrane voltage changes associated with impulse propagation from a narrow cell strand to a large cell area using multiple site optical recording of transmembrane voltage (MSORTV) in conjunction with patterned growth of neonatal rat heart cells in culture. While action potential propagation was smooth in the case of funneled expansions, delays of variable size occurred during propagation into rectangular or incised expansions. Close to the abrupt expansion, which functionally represented an increased electrical load to the narrow cell strand, the delays were accompanied by marked distortions of the action potential upstroke, exhibiting, in extreme cases, an initial depolarization to 50% followed by a delayed secondary depolarization to 100% of the full-signal amplitude. These distortions, which were based on bidirectional electrotonic interactions across the transition, were maximal immediately downstream from the expansion. The maximal slowing of impulse conduction across abrupt expansions was, in agreement with recently published results obtained from two-dimensional computer simulations, always situated in the expanded region. At high stimulation rates, the delays sometimes turned into intermittent unidirectional blocks, as revealed by reverse stimulation. These blocks were always characterized by a marked abbreviation of the action potentials upstream from the region causing the block which might, in an appropriate network, facilitate reentry because of the associated shortening of the refractory period. Because the patterns were composed of cells having identical membrane properties, the results show that the local action potential shape can be modulated profoundly by the two-dimensional architecture of the underlying cell ensemble alone.

Circulation ◽  
2008 ◽  
Vol 118 (suppl_18) ◽  
Author(s):  
David S Auerbach ◽  
Sergey Mironov ◽  
Jose Jalife

Background: Heart diseases predispose to arrhythmias and sudden cardiac death by mechanisms that are poorly understood. We tested the hypothesis that in fully excitable tissue abrupt geometrical expansions resulting from varying wall thickness, replacement fibrosis, ischemia, or accessory pathways, set the stage for to-and-fro propagation (reflection) of electrical impulses over the same pathway, leading to premature excitation and reentry initiation. Methods: We used patterned monolayers of cultured neonatal rat ventricular myocytes consisting of two wide regions connected by a fully excitable, thin isthmus (0.1, 0.5, 1, & 2 mm wide, 6 mm long). We compared control monolayers with those overexpressing Na channels (NaCh, Ad-hSCN5a). Impulse propagation was optically imaged (Di-8-ANEPPS) at high resolution. Results: Impulses initiated proximally in a wide region propagated 1:1 through relatively broad isthmuses (2 mm) and then into the distal expansion at higher frequencies than through narrower isthmuses (0.1 mm). In control monolayers, with relatively low excitability, the prevalence of reflection was small (15%, n=61). NaCh overexpression increased excitability. It also increased the incidence of reflection (38%, n=26). In homogeneous monolayers, NaCh overexpression increased the conduction velocity (15–17%) and prolonged the action potential duration (APD, 21–26%) at the frequencies (2– 4 Hz) at which reflection occurred. During reflection, the APD at the distal expansion was prolonged, compared to APDs within the isthmus or distally when there was no reflection. APD prolongation provided a substrate for local phase-3 re-excitation at the isthmus, and thus reflection. In some cases, reflection was sustained over several consecutive beats. Reflection also triggered the initiation of reentry; as the beat reflected back, a small distal region was re-excited, resulting in unidirectional propagation and the initiation of reentry. Reflection was never observed in structurally homogeneous monolayers, whether with or without the presence of NaCh overexpression. Conclusion: Source-to-sink mismatch in areas of cardiac muscle expansion creates APD heterogeneities, which may serve as a substrate for reflection and arrhythmogenesis.


Circulation ◽  
2008 ◽  
Vol 118 (suppl_18) ◽  
Author(s):  
Gerard J Boink ◽  
Martijn L Bakker ◽  
Arie O Verkerk ◽  
Diane Bakker ◽  
Jacques M de Bakker ◽  
...  

Introduction: Currently constructed biopacemakers based on single gene transfer strategies function suboptimally, with periods of slow heart rates and instabilities during rest. In the sinoatrial node (SAN), the dominant native pacemaker, multiple genes are required for proper impulse formation and impulse propagation. TBX3 is an important regulator of the SAN gene program during heart development. We examined the effects of inducible TBX3 overexpression in adult hearts and in vitro we explored whether lentiviral TBX3 overexpression may be used in biopacemaker engineering. Methods: In vivo atrial and ventricular expression levels of the connexin isoforms Cx43 and Cx40 (impulse propagation) and SCN5A were studied in mice with tamoxifen inducible overexpression of TBX3 using quantitative PCR analysis. Single neonatal rat cardiac myocytes were transduced with TBX3 expressing lentivirus to analyze the effects of TBX3 on action potentials and membrane currents (impulse formation) using the perforated patch-clamp technique. Results: In vivo, Cx43, Cx40 and SCN5A, which are not or only moderately expressed in the native SAN, were severely down-regulated to 20%, 15%, and 40%, respectively, by TBX3 (n=12; p<0.01). Single neonatal cardiac myocytes overexpressing TBX3 exhibited faster spontaneous beating rates, along with decreased maximum diastolic potential, inward rectifier potassium current (I K1 ), and fast sodium current (I Na ). These properties are typical of SAN pacemaker cells. Conclusions: TBX3 can act as a strong repressor of the working myocardium gene program in the adult heart. Overexpression of TBX3 might be a useful tool in biopacemaker gene and cell therapy.


1997 ◽  
Vol 3 (S2) ◽  
pp. 803-804
Author(s):  
B.M. Salzberg ◽  
A.L. Obaid

Molecular indicators of membrane potential may be used to obtain sub-millisecond time resolved images of transient changes in membrane voltage in a variety of biological systems. These probes are small amphipathic molecules having molecular weights of 400-500, and dimensions on the order of 10 Angstroms, which bind to, but do not cross cell membranes, and change either their absorbance or fluorescence in response to membrane voltage. These extrinsic optical signals depend linearly upon membrane potential, and the best of the dyes respond to a step change in voltage in less than 1.5 μsec at room temperature. The salient properties of fast potentiometric probes will be discussed, and the fidelity of optical recordings to transmembrane voltage changes will be considered.Since voltage changes in excitable cells take place on a time scale that is determined by the kinetics of conformational changes in membrane proteins, and by membrane electrical time constants, these changes tend to be very rapid, and resolving them requires imaging systems that are frequently orders of magnitude faster than usual video rates.


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